The Prairie vole resides in dry fields that contain a cover of grasses and weeds, the largest populations are typically found in fallow fields or hay fields.[3] Prairie voles make shallow underground burrows and runways through surface vegetation. In winter, they tunnel underneath the snow. Their runways are used for many purposes, from predator protection to obtaining food. Prairie voles are easily disturbed. They will not hesitate to use their underground burrows if they notice predators close by or disturbances that pose a threat. Compared to the meadow vole, prairie voles prefer to inhabit drier areas.

Prairie voles are active year-round. In colder weather, they tend to be more active during the day; at other times, they are mainly nocturnal. Prairie voles live in colonies and have been known to exhibit human-like social behavior in groups.[4]

Prairie voles live rarely longer than one or two years. Their life expectancy is based on predator presence and natural factors in their area of inhabitance.

During mating season, prairie voles take up individual territories and defend them from other voles. They mark their territories with urine and other secretions. They assume a defensive posture towards a competitor or enemy by raising the forefeet, extending the head forward, and chattering of the teeth. Outside the mating seasons, the prairie voles live together.

Like other voles, prairie voles can reproduce at any time of the year, but the main breeding seasons are in the fall and the spring. Unlike other voles, prairie voles are generally monogamous. The prairie vole is a notable animal model for studying monogamous behavior and social bonding because male and female partners form lifelong pair bonds, huddle and groom each other, share nesting and pup-raising responsibilities, and generally show a high level of affiliative behavior. However, they are not sexually faithful, and though pair-bonded females usually show aggression toward unfamiliar males, both sexes will occasionally mate with other voles if the opportunity arises.[5]

The female's gestation period is between 20 and 30 days. Female voles have two to four litters of two to seven young per year in a nest lined with vegetation in an underground burrow or in a depression on the ground. Litter size varies depending on food availability and the age of the female. The largest number of pregnancies with the highest offspring occur in spring and fall.[3] Baby voles open their eyes at about eight days after birth, and become capable of feeding themselves at about two weeks.

Prairie voles are important to the ecosystem. They provide food for predators, but are considered pests by some. Many ways to prevent voles from destroying gardens or other areas are available. Electric repellers and predators (snakes, owls, coyotes, foxes, domestic animals, and hawks) can be used to reduce vole populations. They can also be scared away by plastic ornaments that resemble natural predators.

Though poison is an option to prevent voles, poisoned voles can create a threat to other animals and humans. Voles are prey for other predators. If they are eaten by predators while poisoned, the poison could harm the predator. In addition, when placing poison near vole entrances, other animals may be able to reach it, making it a hazard to them. Moreover, poison left in the field can easily be blown or washed away. In residential areas, the poison itself and poisoned voles can be harmful and/or dangerous to people and domesticated animals. If a licensed pest control company is involved they can mitigate any poisoning concerns through the use of proper exterior bait stations. .

The prairie vole is special for having pair bonding with its partner.[6] The male prairie vole has a continuous contact with its female counterpart, which lasts for all of their lives. If the female prairie vole dies, the male does not look for a new partner. Moreover, this constant relationship is more social than sexual. For this pair bonding to take place, the male must stay one day with the female after they breed. Other species, such as the montane vole, do not show this pair bonding behavior.

This uniqueness in the prairie vole behavior is related to the oxytocin and vasopressinhormones. The oxytocin receptors of the female prairie vole brain are located more densely in the reward system, and have more receptors than other species, which causes 'addiction' to the social behavior. In the male prairie vole, the gene for the vasopressin receptor has a longer segment, as opposed to the montane vole, which has a smaller segment. This segment is longer in other bonding animals (such as humans[citation needed]), and shorter in other nonbonding animals (such as chimpanzees).